Integrated Analysis of Small RNA, Transcriptome, and Degradome Sequencing Reveals the MiR156, MiR5488 and MiR399 are Involved in the Regulation of Male Sterility in PTGMS Rice.

A photoperiod- and thermo-sensitive genic male sterile (PTGMS) line is the basic material for two-hybrid rice and is an important genetic breeding resource. Peiai64S (PA64S) is an important germplasm resource of PTGMS rice, and it has been applied to two-line hybrid rice systems in China. Pollen fertility in PA64S is regulated by the temperature and photoperiod, but the mechanism of the fertility transition is unclear. In this study, we obtained the male fertile plant PA64S(F) and the male sterile plant PA64S(S) by controlling different temperatures under long light conditions and used the male fertile and sterile plants to investigate the role of microRNAs (miRNAs) in regulating male fertility in rice. We performed the small RNA library sequencing of anthers from PA64S(S) and PA64S(F). A total of 196 miRNAs were identified-166 known miRNAs among 27 miRNA families and 30 novel miRNAs. In the transcriptome analysis, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of differentially expressed genes revealed significant enrichment in the synthesis and metabolism of fatty acids and some secondary metabolism pathways such as fatty acid metabolism and phenylalanine metabolism. With a comprehensive analysis of miRNA, transcriptome, and degradome sequencing, we identified that 13 pairs of miRNA/target genes regulated male fertility in rice by responding to temperature change, among which the miR156, miR5488, and miR399 affect the male fertility of PA64S by influencing SPLs, the lignin synthesis of anther walls, and the flavonoid metabolism pathway. The results provide a new understanding of PTGMS rice, which will help us better understand the potential regulatory mechanisms of male sterility in the future.

Development and validation of a nomogram for predicting survival in patients with resected non-small-cell lung cancer.

PURPOSE: A nomogram is a useful and convenient tool for individualized cancer prognoses. We sought to develop a clinical nomogram for predicting survival of patients with resected non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: On the basis of data from a multi-institutional registry of 6,111 patients with resected NSCLC in China, we identified and integrated significant prognostic factors for survival to build a nomogram. The model was subjected to bootstrap internal validation and to external validation with a separate cohort of 2,148 patients from the International Association for the Study of Lung Cancer (IASLC) database. The predictive accuracy and discriminative ability were measured by concordance index (C-index) and risk group stratification. RESULTS: A total of 5,261 patients were included for analysis. Six independent prognostic factors were identified and entered into the nomogram. The calibration curves for probability of 1-, 3-, and 5-year overall survival (OS) showed optimal agreement between nomogram prediction and actual observation. The C-index of the nomogram was higher than that of the seventh edition American Joint Committee on Cancer TNM staging system for predicting OS (primary cohort, 0.71 v 0.68, respectively; P < .01; IASLC cohort, 0.67 v 0.64, respectively; P = .06). The stratification into different risk groups allowed significant distinction between survival curves within respective TNM categories. CONCLUSION: We established and validated a novel nomogram that can provide individual prediction of OS for patients with resected NSCLC. This practical prognostic model may help clinicians in decision making and design of clinical studies.

Long-term survival outcomes of video-assisted thoracic surgery for patients with non-small cell lung cancer.

BACKGROUND: Video-assisted thoracic surgery (VATS) has been shown to be a safe alternative to conventional thoracotomy for patients with non-small cell lung cancer (NSCLC). However, popularization of this relatively novel technique has been slow, partly due to concerns about its long-term outcomes. The present study aimed to evaluate the long-term survival outcomes of patients with NSCLC after VATS, and to determine the significant prognostic factors on overall survival. METHODS: Consecutive patients diagnosed with NSCLC referred to one institution for VATS were identified from a central database. Patients were treated by either complete-VATS or assisted-VATS, as described in previous studies. A number of baseline patient characteristics, clinicopathologic data and treatment-related factors were analyzed as potential prognostic factors on overall survival. RESULTS: Between January 2000 and December 2007, 1,139 patients with NSCLC who underwent VATS and fulfilled a set of predetermined inclusion criteria were included for analysis. The median age of the entire group was 60 years, with 791 male patients (69%). The median 5-year overall survival for Stage I, II, III and IV disease according to the recently updated TNM classification system were 72.2%, 47.5%, 29.8% and 28.6%, respectively. Female gender, TNM stage, pT status, and type of resection were found to be significant prognostic factors on multivariate analysis. CONCLUSIONS: VATS offers a viable alternative to conventional open thoracotomy for selected patients with clinically resectable NSCLC.

Thoracoscopic half carina resection and bronchial sleeve resection for central lung cancer.

BACKGROUND: The objectives of this study were to report the surgical techniques and clinical outcome of thoracoscopic half carina resection and thoracoscopic bronchial sleeve resection for central lung cancer. METHODS: Between January 2011 and November 2012, 675 patients with lung cancer underwent radical surgery by thoracoscopy, and 49 (7.3%) underwent bronchial sleeve resection. Among 49 patients, 20 (41%) received thoracoscopic bronchial sleeve lobectomy. Perioperative variables and postoperative outcomes of these cases were analyzed to evaluate the technical feasibility and safety of this operation. RESULTS: In one patient, right upper lung sleeve resection was combined with half-carinal resection and reconstruction. In another, right medial lung sleeve resection was combined with lower right dorsal segment resection. The average time of surgery was 239 ± 51 minutes (range = 142-330 minutes), and the average time of airway reconstruction was 44 ± 17 minutes (range = 22-75 minutes). The intraoperative blood loss averaged 207 ± 96 mL (range = 80-550 mL). The median postoperative hospital stay was 10 days (interquartile range = 8-12 days). Postoperatively, extubation was achieved in the recovery room without further need for mechanical ventilation. None of the patients developed anastomotic leak. Perioperative mortality was not observed. CONCLUSION: Thoracoscopic bronchial sleeve resection can be considered a feasible and safe operation for selected patients with central lung cancer. The complicated anastomosis technique of half carina resection was feasible.

Video-assisted thoracoscopic lobectomy for non-small cell lung cancer in patients with severe chronic obstructive pulmonary disease.

OBJECTIVE: To assess the feasibility, safety and long-term outcomes of video-assisted thoracic surgery (VATS) lobectomy for the treatment of non-small cell lung cancer (NSCLC) in patients with severe chronic obstructive pulmonary disease (COPD). METHODS: The clinical data of patients with NSCLC and severe COPD (preoperative FEV1% <50%) who underwent VATS lobectomy from January 2000 to January 2011 were retrospectively analyzed to identify their demographic parameters, postoperative complications and outcomes. RESULTS: The preoperative FEV1/FVC was <70% and FEV1% <50% in all 61 patients in this study, with a mean preoperative FEV1 of 0.99 L (0.54-1.58 L) and mean FEV1% of 38.4% (22-49.82%). All of the 61 patients underwent the VATS lobectomy or sleeve resection plus systemic lymph node dissection. The mean operative time was 218 minutes (120-355 minutes), with a mean intraoperative blood loss of 342 mL (50-1,600 mL). None of the patients converted to thoracotomy. Multivariate statistical analysis revealed that age and TNM staging after tumor resection were independent predictive factors for the 5-year survival in those patients (P=0.014 and 0.013). CONCLUSIONS: With preoperative imaging studies, pulmonary function assessment and target positioning, VATS lobectomy can be safely and effectively performed for patients with NSCLC and severe COPD to achieve a satisfying long-term survival outcome.

Feasibility of complete video-assisted thoracoscopic surgery following neoadjuvant therapy for locally advanced non-small cell lung cancer.

OBJECTIVE: To explore the feasibility of complete video-assisted thoracoscopic surgery (c-VATS) following neoadjuvant therapy (chemotherapy, targeted therapy and radiotherapy, either alone or in combination) for the treatment of patients with non-small cell lung cancer (NSCLC). METHODS: The clinical data of 43 NSCLC patients undergoing c-VATS following neoadjuvant therapy were retrospectively analyzed, including the preoperative functional indicators, staging, concurrent diseases, surgical techniques, operation time, number of lymph nodes dissected and postoperative drainage time and quantity, postoperative hospital stay, postoperative complications, and survival. RESULTS: From January 2006 to March 2012, a total of 43 patients with stage IIA-IIIB NSCLC were included in this study (IIIA: 27 cases, 62.8%; IIIB: 11 cases, 25.6%), including 32 males (74.4%) and 11 females (25.6%). Forty-two patients were operated successfully, 28 underwent pulmonary lobectomies (including 9 bronchial sleeve resections), 5 had double lobectomies, 5 had wedge resections, and 4 had total pneumonectomies. Seven patients were referred to undergo Hybrid VATS (7/42, 16.7%). The mean length of the operation was 160.48±16.52 min (range, 130-180 min); the intraoperative blood loss was 253.57±117.08 mL; the number of lymph nodes dissected was 16.88±10.93; the postoperative drainage time was 1-7 d (mean: 2.62±0.96 d); and the postoperative hospital stay was 3-7 d (mean: 5.45±1.30 d). The incidence of postoperative complications was 9.5% (4/42), and the perioperative mortality was 2.4% (1/42). The 1-, 2-, and 3-year overall survival rates were 94%, 79%, and 65%, respectively. CONCLUSIONS: c-VATS following neoadjuvant therapy is safe and feasible for the treatment of locally advanced NSCLC.

High expression levels of class III ß-tubulin in resected non-small cell lung cancer patients are predictive of improved patient survival after vinorelbine-based adjuvant chemotherapy.

The aim of the present study was to determine the frequency and predictive value of the expression of tumor microtubule components in patients with resected non-small cell lung cancer (R-NSCLC) subsequently treated with vinorelbine-based adjuvant chemotherapy. The expression of the microtubule components was evaluated in 85 R-NSCLC tumor samples using immunohistochemistry. All patients received vinorelbine-based chemotherapy. The predictive value of microtubule protein expression for disease-free survival (DFS) and overall survival (OS) was assessed. The expression of the microtubule components was not associated with any baseline clinicopathological factors in the R-NSCLC patients. High tumor expression levels of class III ß-tubulin were correlated with an improved DFS (P=0.033) and a trend towards a longer OS (P=0.226). Class II and IV ß-tubulins were not correlated with patient outcome. Multivariate analysis of factors, including gender, age, histology, stage and class II, III and IV ß-tubulin expression demonstrated that high levels of class III ß-tubulin expression were correlated independently with DFS (P= 0.031). These findings suggest that high class III ß-tubulin expression levels in resected tumors are predictive of improved DFS in R-NSCLC patients receiving vinorelbine-based chemotherapy.

Video-assisted thoracoscopic surgery for stage I non-small cell lung cancer: long-term survival and prognostic factors.

The purpose of this study was to evaluate the long-term outcomes of video-assisted thoracoscopic surgery (VATS) major pulmonary resection in patients with stage I non-small cell lung cancer (NSCLC). Of the 411 stage I patients, 318 (77.4 %) underwent complete VATS (c-VATS), while 89 (21.7 %) underwent assisted VATS (a-VATS). There were no intraoperative deaths. There were three deaths (0.7 %) within 30 postoperative days. The 1-, 3-, and 5-year survival rates were 95.1 % (95 % CI, 92.9­97.3 %), 83.1 % (95 % CI, 79.2­87.0 %), and 73.4 % (95 % CI, 68.1­78.7 %), respectively. Univariate analysis by log-rank test revealed that tumor-node-metastasis (TNM) stage, primary tumor (pT) status, and type of resection were statistically significant factors affecting overall survival (OS; P = 0.029, P = 0.025, and P = 0.005, respectively). Less acute TNM stage and less extensive resection were significantly predictive for longer OS by multivariate analysis as well (P = 0.024 and P = 0.006, respectively). In experienced hands, c-VATS or a-VATS can be considered as an alternative to traditional incision in patients with stage I NSCLC. Lower TNM stage and less extensive resection were significantly predictive for better OS. A prospective randomized controlled study on a larger scale is required to reach definitive conclusions regarding the efficacy of VATS relative to other techniques.

[EGFR gene copy number, ERCC1 and BRCA1 protein expression and their relationship in non-small cell lung cancer].

OBJECTIVE: To evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them. METHODS: The status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed. RESULTS: FISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P = 0.048), and non-smoker than smoker (32.8% vs. 16.7%, P = 0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P > 0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P = 0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P > 0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P > 0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r = 0.449, P < 0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein. CONCLUSIONS: FISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.

Prognostic impact of MMP-2 and MMP-9 expression in pathologic stage IA non-small cell lung cancer.

BACKGROUND: The purpose of the present study was to assess the value of matrix metalloproteinase (MMP)-2 and MMP-9 expression and other potential prognostic factors in predicting the clinical outcome of patients after definitive surgery for pathologic stage IA non-small cell lung cancer (NSCLC). METHODS: One hundred and forty-six consecutive and non-selected patients who underwent definitive surgery for stage IA NSCLC were included in this study. Formalin-fixed paraffin-embedded specimens were stained for MMP-2 and MMP-9, which were statistically evaluated for their prognostic value and other clinicopathological parameters. RESULTS: Of the 146 patients studied, 102 (69.9%) cases were classified as having high expression for MMP-2. A total of 89 carcinomas (61.0%) had high expression for MMP-9. MMP-9 expression correlated with Eastern Cooperative Oncology Group (ECOG) performance status, pT stage, and differentiation (P = 0.005, <0.001, and <0.001, respectively). Vessel invasion, pT stage, and MMP-9 expression maintained their independent prognostic influence on overall survival (P = 0.037, <0.001, and <0.001, respectively). CONCLUSIONS: From results of our relatively large database, MMP-9 may be considered as a viable biomarker that can be used in conjunction with other prognostic factors such as vessel invasion and pT stage to predict the prognosis of patients with completely resected pathologic stage IA NSCLC.

Long-term outcome and cost-effectiveness of complete versus assisted video-assisted thoracic surgery for non-small cell lung cancer.

BACKGROUND: To compare the outcomes and costs of two methods of video-assisted thoracoscopic surgery (VATS) major pulmonary resection in patients with clinically resectable non-small cell lung cancer (NSCLC). METHODS: Between January 2000 and December 2007, 1,058 patients with proven stages I-IIIA NSCLC underwent complete VATS (c-VATS) or assisted VATS (a-VATS) major pulmonary resection together with a systematic nodal dissection. RESULTS: The study cohort consisted of 736 men and 322 women. Mean operative time was shorter for the a-VATS cohort compared with the c-VATS group (P = 0.038). Overall survival (OS) at 5 years based on Kaplan-Meier analysis was 55.3% (95%CI, 50.6-60.0%) for those who underwent c-VATS and 47.7% (95%CI, 41.2-54.2%) for those who underwent a-VATS (P = 0.404). Gender, final pathology, TNM stage, and pT status were significant predictive factors for OS according to multivariate analysis. The total cost of a-VATS lobectomy was lower than that of c-VATS lobectomy. CONCLUSIONS: c-VATS and a-VATS yield similar results in patients with clinically resectable NSCLC. a-VATS, however, may be less expensive and easier to adopt, making it a particularly attractive option for thoracic surgeons in developing countries.

Long-term outcome of hybrid surgical approach of video-assisted minithoracotomy sleeve lobectomy for non-small-cell lung cancer.

BACKGROUND: The aim of this study was to evaluate the technical feasibility and safety of a hybrid surgical approach of video-assisted minithoracotomy (hybrid VATS) sleeve lobectomy for non-small-cell lung cancer (NSCLC), using success rate as the primary end point. METHODS: Between February 1996 and December 2006, patients with bronchogenic tumors were prospectively registered to undergo hybrid VATS sleeve resection in a single institution. Hybrid VATS involved performing the main procedures via rib spreading and minithoracotomy using a monitor and direct vision. A successful procedure was defined as a patient who had a sleeve lobectomy via hybrid VATS without conversion to thoracotomy and without significant perioperative morbidity or mortality. RESULTS: A total of 148 patients (108 men and 40 women; median age = 58 years) who underwent hybrid VATS sleeve lobectomy for NSCLC were identified in our database. The median duration of the successfully completed procedures was 190 min (range = 145-305 min). The median length of time of chest tube in place was 3 days (range = 1-12 days). Hybrid VATS sleeve lobectomy was performed successfully in 134 of 148 patients for a success rate of 90.5%. The median follow-up period was 65.1 months (range = 34.5-154.8 months). The overall 5-year disease-free survival and overall survival of all patients were 36.7% (95% CI = 27.9-45.5%) and 54.2% (95% CI = 44.8-63.6%), respectively. CONCLUSION: Hybrid VATS sleeve lobectomy is feasible for selected patients with NSCLC in specialized centers.

[Video-assisted mini-thoracoscopy for complete resection of 28 cases of pleomorphic carcinoma of the lung].

BACKGROUND AND OBJECTIVE: Video-assisted mini-thoracoscopy (VAMT) has been used for pulmonary lobectomy for 20 years, which has many merits including small wound, less pain and quick recovery. VAMT is the tendency of pectoral minimally invasive surgical treatment. The aim of this study is to evaluate the efficacy of video-assisted mini-thoracoscopy (VAMT) for radical resection of pleomorphic carcinoma of the lung. METHODS: Complete resection of pleomorphic carcinoma of the lung was performed on 28 patients by VMAT. A 1.5 cm and a 6 cm to 8 cm incision was made during the operation. Both standard surgical instruments and thoracoscopic set were used to treat the pulmonary vessels, perform lobotomy, and remove the lymph nodes in the mediastinum and pulmonary portal. RESULTS: The operation was completed in all of the cases. No peri-operative death occurred. The total volume of hemorrhage was 200 mL to 450 mL (mean, 300 mL). The patients received chest drainage for 3 to 8 days after the operation (mean, 5 d). The time in hospital was 7 d-14 d (mean, 12 d). Five-year overall survival and disease-free survival were 39.2% and 47.1%, respectively. Follow-up was available in all 28 patients for up to 2 to 91 months. Among the 28 cases, only 2 patients died 2 months after the operation, the rest all live up over 1 year. 3-year survival rate was 60.7% (95%CI: 40.3%-81.1%). CONCLUSION: VAMT is effective for radical resection of pleomorphic carcinoma of the lung in a short term. Combination the superiority of traditional procedure and VATS, so VAMT is safe and reliable for radical resection of pleomorphic carcinoma of the lung.

[Correlation between pre-treatment anemia and prognosis in non-small cell lung cancer patients].

BACKGROUND AND OBJECTIVE: The patients with non-small cell lung cancer (NSCLC) might contract anemia, however, whether anemia is one of the independent prognostic factors to the patients with NSCLC is still controversial. So the aim of this study is to investigate the correlation between anemia and overall survival (OS) in patients with NSCLC. METHODS: 1 018 patients with operable NSCLC were retrospectively analyzed in our hospital from January 2000 to December 2008. RESULTS: The occurrence of anemia before operation was 252/1 018 (24.1%). The OS in NSCLC patients without anemia was (2 425.98 +/- 50.03) days, and the OS in patients with anemia was (2 107.15 +/-93.86) days. There was significant difference in the OS between them (P = 0.001). The patients with anemia in stage I had shorter survival time than those without anemia (P < 0.001). But there was no difference in other stage patients. TNM stage, gender, tumor size and lymph nodes metastasis were correlated with OS using Cox regression analysis. CONCLUSIONS: Anemia is correlated with survival in operable NSCLC patients. Moreover, it is an independent prognostic factor in NSCLC patients with stage I.

Detection of CEA mRNA, p53 and AE1/AE3 in haematoxylin-eosin-negative lymph nodes of early-stage non-small cell lung cancer may improve veracity of N staging and indicate prognosis.

OBJECTIVE: Although the surgical-pathological classification can be considered the 'gold standard' of T-N staging, it could not provide satisfactory and accurate estimation of survival rates in early-stage non-small cell lung cancer (NSCLC). METHODS: In our study, the expression of carcinoembryonic antigen (CEA), p53 and intracytoplasmic keratin (AE1/AE3) using haematoxylin-eosin (HE) staining negative lymph nodes (LNs) in 28 patients with early-stage NSCLC were analysed using fluorescent quantitation reverse transcription-polymerase chain reaction (FQ-PCR) and immunohistochemistry (IHC). RESULTS: One hundred and ninety-three LNs were analysed. Two patients staged as I up-staged to II, and six patients staged as II up-staged to III. About 32, 19 and 36 LNs were positive, respectively, for CEA mRNA (32/193, 16.6%), p53 (19/193, 9.84%) and AE1/AE3 (36/193, 18.65%) compared with control LNs. Only FQ-PCR test for CEA mRNA could detect micrometastases in stage I NSCLC patients with N0 LNs (2/13, 15.4%). Disease-free time in patients with CEA mRNA (P = 0.000), p53 protein (P = 0.013) and AE1/AE3 (P = 0.003) positive were significantly inferior to those with micrometastases negative. Moreover, the results demonstrated that the positive LNs for CEA mRNA (P = 0.028), p53 protein (P = 0.048) and AE1/AE3 (P = 0.007) were associated with the relapse time, respectively. However, Cox proportional hazards test showed that only clinical stage was the independent risk factor of relapse, and denied the correlation between micrometastases in LNs and recurrence. CONCLUSIONS: Detection of CEA mRNA, p53, AE1/AE3 in HE-negative LNs may improve veracity of N staging and predict its prognosis in patients with early-stage NSCLC. Furthermore, micrometastases in stage I may be performed by FQ-PCR more sensitive than IHC.

[Treatment and timing of re-operation for postoperative recurrence of spontaneous pneumothorax].

OBJECTIVE: To investigate the treatment for postoperative recurrence of spontaneous pneumothorax, the feasibility of re-operation, indications and the results. METHOD: The clinical and follow-up data of 28 cases of postoperative recurrence from 485 patients who underwent operations for spontaneous pneumothorax from Jan, 1994 to Jan, 2005 were retrospectively reviewed. RESULTS: One case was observed without any invasive therapy, 12 cases underwent closed pleural drainage and 15 cases received reoperation. All of the patients had the lungs reexpanded sufficiently, with comorbidities in 2 cases, one of which was pleural effusion, the other was prolonged leakage after surgery. No death occurred. The operation time was 80 - 315 (115 +/- 43) min, and the bleeding volume was 50 - 350 (115 +/- 54) ml. The time of chest tube drainage was 1 - 7 (4 +/- 2) d, and the time of hospital stay was 7 - 18 (11 +/- 5) d. The follow-up of the 28 cases ranged from 1 to 107 months (median 55.4) without recurrence. CONCLUSIONS: The findings on lung high rate CT (HRCT) are the most important factors in deciding treatments of postoperative recurrence of pneumothorax. If it is the first recurrence, and there is no evident bullae on HRCT, conservative therapy should be considered, especially for the recurrence within two years postoperation. If the first recurrence occurs after surgery, especially within 2 years, and the bullae is invisible on HRCT, adhesion is the choice without significant adverse effect. If the recurrence occurred twice and more, bullae is found on HRCT, adhesion hampered lung reexpansion, or pleural drainage alone could not work, re-operation should be performed.